Dataset Information

Practical Protocol for Making Calibration Curves for Direct and Sensitive Quantitative LC Orbitrap-MS of Large Neuropeptides.

ABSTRACT: Peptides larger than 3-4?kDa, such as neuropeptide Y (NPY), orexin-B, and alpha-MSH, have practical issues that arise when conducting direct and sensitive quantitative liquid chromatography (LC) orbitrap-FT mass spectrometry (MS) due to their adsorption and low ionization efficiency, especially in standard solutions. A mixing solvent consisting of 0.5% trifluoroacetic acid (TFA) and 35-50% aq. acetonitrile was developed as the standard NPY for creating calibration curves, as well as a matrix to block the experimental tube surface to minimize adsorption. The mixture matrix effectively blocked non-specific adsorption of the standard peptides with tryptic digested bovine serum albumin (BSA) (small fragment peptides) and orexin-B (a large chain peptide). A sample containing 1?:?100 peptide:water was detected in the developed sample solution. Finally, 2 to 1,000?fmol/?L NPY could be analyzed quantitatively and reproducibly using conventional LC-MS. Parameters of the calibration curves, such as X-intercept, Bias (%), and relative standard deviation (RSD), were adjusted to optimize the sample solutions and the sensitive and quantitative LC-MS analyses.

SUBMITTER: Yamagaki T

PROVIDER: S-EPMC7392841 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Practical Protocol for Making Calibration Curves for Direct and Sensitive Quantitative LC Orbitrap-MS of Large Neuropeptides.

Yamagaki Tohru T Yamazaki Takashi T

Mass spectrometry (Tokyo, Japan) 20200803 1

Peptides larger than 3-4 kDa, such as neuropeptide Y (NPY), orexin-B, and alpha-MSH, have practical issues that arise when conducting direct and sensitive quantitative liquid chromatography (LC) orbitrap-FT mass spectrometry (MS) due to their adsorption and low ionization efficiency, especially in standard solutions. A mixing solvent consisting of 0.5% trifluoroacetic acid (TFA) and 35-50% aq. acetonitrile was developed as the standard NPY for creating calibration curves, as well as a matrix to ...[more]

PMID: 32802701

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Project description:Edible vegetable oils comprise integral components of humans' daily diet during the lifetime. Therefore, they constitute a central part of dietary-exposome, which among other factors regulates human health. In particular, the regular consumption of olive oil (OO) has been largely accepted as a healthy dietary pattern. Responsible for its recognition as a superior edible oil is its exceptional aroma and flavor. Its unique composition is characterized by high levels of monounsaturated fatty acids and the presence of minor constituents with important biological properties, such as the so-called OO polyphenols. Being a high added value product, OO suffers from extensive fraud and adulteration phenomena. However, its great chemical complexity, variability, and the plethora of parameters affecting OO composition hamper significantly the selection of the absolute criteria defining quality and authenticity, and a reliable and robust methodology is still unavailable. In the current study, Flow Injection Analysis-Magnetic Resonance Mass Spectrometry (FIA-MRMS) was investigated under a metabolic profiling concept for the analysis of Greek Extra Virgin Olive Oils (EVOO). More than 200 monovarietal (Koroneiki) EVOO samples were collected from the main Greek OO producing regions and investigated. Both intact oil and the corresponding polyphenols were analyzed in fast analysis time of 2 and 8 min, respectively. In parallel, an LC-Orbitrap MS platform was used to verify the efficiency of the method as well as a tool to increase the identification confidence of the proposed markers. Based on the results, with FIA-MRMS, comparable and improved projection and prediction models were generated in comparison to those of the more established LC-MS methodology. With FIA-MRMS more statistically significant compounds and chemical classes were identified as quality and authenticity markers, associated with specific parameters, i.e. geographical region, cultivation practice, and production procedure. Furthermore, it was possible to monitor both lipophilic and hydrophilic compounds with a single analysis. To our knowledge, this approach is among the few studies in which two FT-MS platforms combining LC and FIA methods were integrated to provide solutions to quality control aspects of OO. Moreover, both lipophilic and hydrophilic components are analyzed together, providing a holistic quality control workflow for OO.

Dataset Information

Practical Protocol for Making Calibration Curves for Direct and Sensitive Quantitative LC Orbitrap-MS of Large Neuropeptides.

Publications

Practical Protocol for Making Calibration Curves for Direct and Sensitive Quantitative LC Orbitrap-MS of Large Neuropeptides.

Similar Datasets

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